AI Article Synopsis
- This chapter outlines an optimized protocol for isolating synaptic vesicles (SVs) from human induced pluripotent stem cell-derived neurospheres, beginning with the cultivation of mature neurons for functional studies.
- The process involves isolating neurosphere-derived synaptosomes and enriching SVs through differential centrifugation.
- Finally, the protocol utilizes nanoLC-MS/MS for proteomic analysis of SVs, aiding in the understanding of SV molecular diversity and neurotransmitter dynamics, with potential applications in neurological and neuropsychiatric research.
Article Abstract
This chapter presents an optimized method for isolating synaptic vesicles (SVs) from neurospheres derived from human induced pluripotent stem cells (hiPSCs). The protocol begins with neurosphere cultivation to achieve mature neurons, which is essential for the functional studies of neuronal activity. Following this, neurosphere-derived synaptosomes are isolated, and SVs are enriched through differential centrifugation. The method culminates in the proteomic analysis of SVs using nano-liquid chromatography coupled with high-resolution tandem mass spectrometry (nanoLC-MS/MS), providing a detailed proteome profile of the isolated vesicles. This protocol can contribute to the understanding of SV molecular heterogeneity and the mechanisms of neurotransmitter uptake and release and be applied to the field of research in neurological and neuropsychiatric disorders.
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| http://dx.doi.org/10.1007/978-1-0716-4298-6_14 | DOI Listing |
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